Hyaline Cartilage Microtissues Engineered from Adult Dedifferentiated Chondrocytes: Safety and Role of WNT Signaling.

The repair of damaged articular cartilage is an unmet medical need. Chondrocyte-based cell therapy has been used to repair cartilage for over 20 years despite current limitations. Chondrocyte dedifferentiation upon expansion in monolayer is well known and is the main obstacle to their use as cell source for cartilage repair. Consequently, current approaches often lead to fibrocartilage, which is biomechanically different from hyaline cartilage and not effective as a long-lasting treatment. Here, we describe an innovative 3-step method to engineer hyaline-like cartilage microtissues, named Cartibeads, from high passage dedifferentiated chondrocytes. We show that WNT5A/5B/7B genes were highly expressed in dedifferentiated chondrocytes and that a decrease of the WNT signaling pathway was instrumental for full re-differentiation of chondrocytes, enabling production of hyaline matrix instead of fibrocartilage matrix. Cartibeads showed hyaline-like characteristics based on GAG quantity and type II collagen expression independently of donor age and cartilage quality. In vivo, Cartibeads were not tumorigenic when transplanted into SCID mice. This simple 3-step method allowed a standardized production of hyaline-like cartilage microtissues from a small cartilage sample, making Cartibeads a promising candidate for the treatment of cartilage lesions.

Macropinocytosis requires Gal-3 in a subset of patient-derived glioblastoma stem cells.

Recently, we involved the carbohydrate-binding protein Galectin-3 (Gal-3) as a druggable target for KRAS-mutant-addicted lung and pancreatic cancers. Here, using glioblastoma patient-derived stem cells (GSCs), we identify and characterize a subset of Gal-3high glioblastoma (GBM) tumors mainly within the mesenchymal subtype that are addicted to Gal-3-mediated macropinocytosis. Using both genetic and pharmacologic inhibition of Gal-3, we showed a significant decrease of GSC macropinocytosis activity, cell survival and invasion, in vitro and in vivo. Mechanistically, we demonstrate that Gal-3 binds to RAB10, a member of the RAS superfamily of small GTPases, and ß1 integrin, which are both required for macropinocytosis activity and cell survival. Finally, by defining a Gal-3/macropinocytosis molecular signature, we could predict sensitivity to this dependency pathway and provide proof-of-principle for innovative therapeutic strategies to exploit this Achilles' heel for a significant and unique subset of GBM patients.

Evaluation of two computational models of amplitude modulation coding in the inferior colliculus.

Two computational models replicating amplitude-modulation encoding in the inferior colliculus (IC) are presented and compared. Neurons in this nucleus are modeled as point neurons using Mc Gregor equations, and receive depolarizing currents from action potentials delivered by stellate cells (chopper units) in the cochlear nucleus (CN). Stellate cells are modeled using modified Hodgkin-Huxley equations and receive inputs from a peripheral auditory model. The CN models of the two proposed architectures are characterized by an important dispersion of cellular characteristics, and therefore by various cellular best modulation frequencies (BMFs) ranging from 60 to 300 Hz. In contrast with the previous model proposed by [M.J. Hewitt, R. Meddis, A computer model of amplitude-modulation sensitivity of single units in the inferior colliculus, J. Acoust. Soc. Am. 95 (1994) 2145], each IC cell model receives convergent input from stellate cells with various BMFs. This approach assumes therefore minimal constraints on the model architecture and cell characteristics. The two models differ in terms of the neuronal structure of the IC, composed of 1 or 2 layers of point neurons acting as coincidence detectors. Each model is evaluated using two metrics: mean firing rate and modulation gain. Rate and temporal modulation transfer functions (r-MTFs and t-MTFs, respectively) are simulated and compared with physiological data. Simulations reveal that (i) an important dispersion of BMFs in the CN cells providing input to IC cells yields plausible IC cells responses to AM stimuli, (ii) the 2-layer IC structure yields the best approximation of IC responses measured in vivo.